Homo sapiens (human)

Orthogonal gene control with a catalytically active Cas9

A CRISPR-based tool to knock out and activate genes in the same cell would provide a powerful method to manipulate biological processes. More...

A CRISPR-based tool to knock out and activate genes in the same cell would provide a powerful method to manipulate biological processes. Here we report that sgRNAs with 14-15 bp target sequences and MS2 binding loops can activate gene expression using an active Cas9 nuclease, without inducing DSBs. We use these ‘dead RNAs’ to perform orthogonal gene knockout and transcriptional activation using wild type Cas9 in human cells. We do whole transcriptome RNA-sequencing to characterize the specificity of these deadRNAs. Less...